Oxylipin Signaling in Congenital Heart Disease

项目摘要 先天性心脏病（CHD）是人类最常见的先天性心脏病， 然而，在大多数情况下，CHD的原因是未知的。脂氧合酶衍生的氧脂是一种 复杂且特征不佳的一类高度氧化的多不饱和脂肪酸衍生的生物活性物质 具有关键生物活性的脂质。我们最近发现了氧脂素信号，人类 心血管发育和CHD。这一发现是至关重要的，不仅因为现有的差距， 知识，而且还因为氧脂素具有强烈的饮食依赖性。我们的初步数据显示 脂氧合酶和它们的氧脂素产物显著调节心肌中胚层的形成， 人心脏分化模型，它们的缺失导致小鼠CHD。我们的核心假设是 内源性未表征的氧脂素是心源性中胚层特化的关键调节剂 以及通过与Wnt信号通路的相互作用而影响心脏形态发生。我们的目标是 深入了解心脏发育过程中的脂氧合酶和氧脂素信号通路， 确定它们对CHD机制的贡献。具体目标是：（1）确定 人心源性中胚层特化中氧脂肽信号传导的分子和细胞机制。 (2)研究氧脂素在人类干细胞心脏细胞外基质组织中的作用- 心脏类器官模型。(3)氧脂素信号通路在先天性心脏病中的作用 老鼠的缺陷方法：我们将研究氧脂素在人多能干细胞衍生的 心源性中胚层和3D心脏类器官模型，通过结合LC-MS脂质组学，细胞和 分子生物学和生物化学方法。我们还将采用脂氧合酶敲除小鼠， 氧脂素饮食限制，以研究氧脂素在小鼠胚胎体内心脏发育中的作用。 意义：这项工作通过探索生物学作用解决了知识上的一个主要空白。 氧脂素在心脏发育中的作用及其与冠心病关系的新发现机制。我们 也提供了一种新的潜在联系饮食和先天性心脏缺陷具有重大意义 冠心病的预防。最后，我们提供了发现新疗法的可能性， 可下药的目标和信号网络创新：我们将首次探索关键 脂肪氧合酶、氧脂素和心脏发育信号网络之间的分子机制 人类和小鼠组织。我们还将建立一个新的心脏发育模型， 这些环境脂质与发育分子网络的作用。影响：该项目将 极大地扩展了心脏发育中的氧脂素生物学知识，帮助制定策略， 预防和潜在治疗冠心病，并开发新的工具，用于研究人类生物学中的氧脂素。

Project summary Congenital heart defects (CHD) constitute the most common type of congenital defect in humans, however the causes for CHD in most cases are unknown. Lipoxygenase-derived oxylipins are a complex and poorly characterized class of highly oxidized, polyunsaturated fatty acid-derived bioactive lipids with key biological activities. We recently found a link between oxylipin signaling, human cardiovascular development and CHD. This finding is critical not only because of the existing gap in knowledge, but also because oxylipins are strongly dietary-dependent. Our preliminary data suggests lipoxygenases and their oxylipin products dramatically modulate cardiogenic mesoderm formation in human cardiac differentiation model, and their loss leads to CHD in mice. Our central hypothesis is that endogenous uncharacterized oxylipins are critical regulators of cardiogenic mesoderm specification and heart morphogenesis through cross-talk with the Wnt signaling pathway. Our objective is to provide insight into lipoxygenase and oxylipin signaling pathways during cardiac development and determine their contribution to mechanisms of CHD. Our Specific aims are: (1) Determine the molecular and cellular mechanisms of oxylipin signaling in human cardiogenic mesoderm specification. (2) Investigate the role of oxylipins in cardiac extracellular matrix organization in a human stem cell- derived cardiac organoid model. (3) Evaluate the contribution of oxylipin signaling to congenital heart defects in mice. Approach: We will study oxylipin function in human pluripotent stem cell-derived cardiogenic mesoderm and in a 3D cardiac organoid model by combining LC-MS lipidomics, cell and molecular biology and biochemical methods. We will also employ lipoxygenase knockout mice and oxylipin dietary restriction to study oxylipins in cardiac development in vivo in mouse embryos. Significance: This work addresses a major gap in knowledge by exploring the biological roles of oxylipins in cardiac development, and mechanisms underlying their newly discovered links to CHD. We also offer a novel potential link between diet and congenital heart defects with significant implications for CHD prevention. Finally, we offer the potential to discover new therapies for treatment by identifying druggable targets and signaling networks. Innovation: We will explore for the first time the key molecular mechanisms linking lipoxygenases, oxylipins and cardiac development signaling networks in human and mouse tissues. We will also establish a new model of cardiac development that integrates the role of these environmental lipids with developmental molecular networks. Impact: This project will dramatically expand knowledge of oxylipin biology in cardiac development, help develop strategies to prevent and potentially treat CHD, and develop new tools for the study of oxylipins in human biology.